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Process Design Examples

Complete downstream processing routes for biopharmaceuticals, industrial biotechnology, and gene therapy — from harvest to final product

Biopharmaceuticals

Monoclonal Antibody (mAb) Purification

CHO cells → IgG
Industry platform process: Protein A capture, ion exchange polishing, viral clearance, UF/DF formulation
Yield: 70–85%
Purity: >99%
Steps: 5–7

Recombinant Insulin Purification

E. coli → Insulin (inclusion bodies)
Inclusion body isolation, refolding, enzymatic cleavage, IEX capture, RPC polishing, crystallization
Yield: 60–75%
Purity: >99%
Steps: 6–8

Vaccine Purification

Cell culture → Viral antigen
Cell lysis, clarification, chromatography capture, polishing, sterile filtration, formulation
Yield: 40–70%
Purity: >95%
Steps: 5–8

Lysosomal Enzyme Replacement

CHO cells → Recombinant enzyme
Secreted enzyme recovery, multi-step chromatography, viral clearance, UF/DF for rare disease therapeutics
Yield: 50–70%
Purity: >98%
Steps: 5–7

Gene & Cell Therapy

AAV Vector Purification

HEK293 → AAV serotypes
Cell lysis, nuclease digestion, affinity capture (AAVX), ion exchange polishing, UF/DF for gene therapy
Yield: 30–60%
Purity: >95%
Steps: 5–7

mRNA Purification

In vitro transcription → mRNA
IVT reaction cleanup, DNase digestion, TFF concentration, oligo-dT affinity, AEX polishing for vaccines
Yield: 50–80%
Purity: >95%
Steps: 4–6

Plasmid DNA Purification

E. coli → pDNA
Alkaline lysis, clarification, AEX capture, HIC polishing, UF/DF for gene therapy vectors
Yield: 40–70%
Purity: >97%
Steps: 5–7

Industrial Biotechnology

Bioethanol Production

Yeast fermentation → Fuel ethanol
Cell removal, beer column distillation, rectification to azeotrope, molecular sieve dehydration to >99.5%
Yield: 90–95%
Purity: >99.5%
Steps: 3–5

Citric Acid Production

Aspergillus niger → Citric acid
Mycelial removal, Ca(OH)₂ precipitation, H₂SO₄ acidification, carbon treatment, crystallization
Yield: 70–80%
Purity: >99%
Steps: 5–7

Industrial Enzyme Recovery

Bacillus / fungi → Protease, amylase, etc.
Cell removal, UF concentration, diafiltration, spray drying for detergent and food enzymes
Yield: 80–90%
Purity: 70–90%
Steps: 3–5

Frequently Asked Questions

What is the standard platform process for monoclonal antibody purification?

The mAb platform uses Protein A affinity capture (60–80% yield, removes >99% HCP), followed by CEX or AEX polishing chromatography (removes aggregates and remaining HCPs), viral inactivation steps (low pH hold + nanofiltration), and final UF/DF for formulation. This sequence consistently achieves >99.5% purity and is used for most FDA-approved mAbs.

How many purification steps are typically needed for a biopharmaceutical?

Most biopharmaceuticals require 3–5 downstream processing steps. Small-molecule fermentation products (amino acids, organic acids) often need only 2–3 steps. Recombinant proteins typically need 4–5 steps. Viral vectors and gene therapy products require 5–8 steps due to stringent purity requirements and complex impurity profiles.

What overall yield should I expect from a multi-step purification process?

A 5-step process with 80% yield per step achieves 0.85 = 33% overall yield. Pharmaceutical biologics typically achieve 30–60% overall yield. Small-molecule products can reach 70–90% with optimized crystallization. To maximize overall yield, minimize the number of steps and optimize each step for yield first, then purity.

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